6533b7ddfe1ef96bd127368f

RESEARCH PRODUCT

Dispersion from Cα or NH: 4D experiments for backbone resonance assignment of intrinsically disordered proteins

Helena TossavainenPerttu PermiRiikka IhalinMaarit HellmanSanteri Salovaara

subject

0303 health sciencesChemical substanceChemistryChemical shiftIDPintrinsically disordered proteinresonanssi010402 general chemistryIntrinsically disordered proteinsAggregatibacter actinomycetemcomitans01 natural sciencesBiochemistryResonance (particle physics)bakteerit0104 chemical sciences03 medical and health sciencesCrystallographyBilRIproteiinitNMR-spektroskopiaDispersion (chemistry)Peptide sequenceresonance assignmentSpectroscopy030304 developmental biology

description

AbstractResonance assignment of intrinsically disordered proteins is remarkably challenging due to scant chemical shift dispersion arising from conformational heterogeneity. The challenge is even greater if repeating segments are present in the amino acid sequence. To forward unambiguous resonance assignment of intrinsically disordered proteins, we present iHACANCO, HACACON and (HACA)CONCAHA, three Hα-detected 4D experiments with Cα as an additional dimension. In addition, we present (HACA)CON(CA)NH and (HACA)N(CA)CONH, new 4D Hα-start, HN-detect experiments which have two NH dimensions to enhance peak dispersion in a sequential walk through C′, NH and HN, and provide more accurate NH/HN chemical shifts than those that can be obtained from a crowded 1H, 15N-HSQC spectrum. Application of these 4D experiments is demonstrated using BilRI (165 aa), an outer-membrane intrinsically disordered protein from the opportunistic oral pathogen Aggregatibacter actinomycetemcomitans. BilRI amino acid sequence encompasses three very similar repeats with a 13-residue identical stretch in two of them.

yearjournalcountryeditionlanguage
2020-01-13Journal of Biomolecular NMR
https://doi.org/10.1007/s10858-020-00299-w